Epidemiology of Fusarium Infection and Deoxynivalenol Content in Winter Wheat in the Rhineland, Germany

Details of our long-term research programme concerning the epidemiology of Fusarium spp. and mycotoxin production are summarized. Evaluation of the occurrence of Fusarium spp., mainly on winter wheat (Triticum aestivum), was carried out by investigating Fusarium infection and mycotoxin contamination. Two to 15% of grains were infested during 1995–1998 at three climatologically differing localities of the Rhineland, Germany. Disease progress was accelerated by rainfall during the flowering season. The species most frequently isolated were Fusarium avenaceum, F. poae, F. culmorum and F. graminearum. The mean deoxynivalenol (DON) content varied from 19gkg^–1 (1995) to 310gkg^–1 (1998) and was not always correlated with disease severity. Organic farming systems showed lower rates of infection with ear blight and lower mycotoxin contamination than conventional farming systems.     

Differences in pathogenesis observed among susceptible interactions of carnation with four races of Fusarium oxysporum f.sp. dianthi

Susceptible interactions of Early Sam carnations with races 1,2,4, and 8 ofFusarium oxysporum f. sp.dianthi differed in pathogenesis, both after stem and after root inoculation. Race 1 induced pallescence and withering of leaves. Affected vascular tissue had a uniform pallid to pale brown colour; though heavily colonized, it was not or virtually not degraded. Defence reactions developed only slowly. Race 2 induced yellowing, of the midribs in particular, and withering of leaves. Affected vascular tissue was white with dark brown margins. Colonized tissue was degraded to leave vascular cavities. At lower heights of colonization, many defence reactions developed, which sometimes resulted in localization of the pathogen. Race 4 induced a similar pathogenesis as race 2, except for less intensive defence reactions. Race 8 induced midrib lesions on, and pallescence, withering and necrosis of leaves. Affected vascular tissue had a uniform light brown colour. Degradation of colonized vascular tissues was rare; instead, many defence reactions were observed, even at high heights in the plants.Races 1, 2 and 4 ofF. oxysporum f. sp.dianthi did not induce disease symptoms in Novada carnations, known to be highly resistant to race 2. Stem-inoculated plants localized the infection close to the inoculation site; stems of root-inoculated plants remained unaffected. The localization response also occurred in Early Sam and Novada carnations stem-inoculated withF. oxysporum f. sp.lycopersici.Samenvatting Tussen interacties van Early Sam-anjers met fysio's 1, 2, 4 en 8 vanF. oxysporum f. sp.dianthi werden verschillen in ziekteontwikkeling gevonden na wortel-zowel als stengelinoculatie. Fysio 1 gaf verbleking en verdroging van de bladeren. Aangetast vaatweefsel was gelijkmatig vaal of lichtbruin van kleur, en werd hevig gekoloniseerd, maar vrijwel niet afgebroken. Afweerreacties kwamen slechts traag op gang. Fysio 2 gaf vergeling, in het bijzonder van de hoofdnerven, en verdroging van de bladeren. Aangetast vaatweefsel was wit met donkerbruine randen. Gekoloniseerd weefsel werd afgebroken, hetgeen leidde tot de vorming van holten in het vaatweefsel. In de lagere gekoloniseerde delen traden veel afweerreacties op, hetgeen soms lokalisatie van het pathogeen tot gevolg had. Fysio 4 gaf eenzelfde ziekteontwikkeling als fysio 2, maar minder afweerreacties. Fysio 8 gaf lesies bij de hoofdnerven, en verbleking, verdroging en necrose van bladeren. Aangetast vaatweefsel was gelijkmatig lichtbruin van kleur. Afbraak van gekoloniseerd vaatweefsel werd zelden waargenomen; veel afweerreacties vergezelden de kolonisatie tot hoog in de stengel.Inoculatie van Novada anjers met fysio's 1,2 en 4 vanF. oxysporum f. sp.dianthi had geen ziektesymptomen tot gevolg. Via de stengel geïnoculeerde planten lokaliseerden de infectie ter hoogte van het inoculatiepunt; de stengels van via de wortels geïnoculeerde planten waren onaangetast. De lokalisatiereactie trad ook op in Early Sam en Novada anjers na inoculatie via de stengel metf. oxysporum f. sp.lycopersici.     

Effect of Fungicides on Fusarium Head Blight and Deoxynivalenol Content in Durum Wheat Grain

In 1998–99 and 1999–2000 six trials were conducted to evaluate the effect of fungicides on Fusarium head blight in the field, on infected kernels and deoxynivalenol (DON) concentration in grain. A single application of prochloraz, tebuconazole, epoxiconazole or bromuconazole, applied to durum wheat varieties at the manufacturer's recommended dose at the beginning of anthesis stage, provided good control of the disease when infective pressure in the field was low to medium, and when the main pathogens were F. graminearum and F. culmorum. Kresoxim-methyl showed a low efficacy at controlling the disease. Tebuconazole, prochloraz and bromuconazole were effective at controlling F. graminearum and F. culmorum, while kresoxim-methyl was not effective in reducing Fusarium infected kernels. DON concentration in grain of cultivars inoculated with F. graminearum and F. culmorum was high, averaging 4.2mgkg^–1 (untreated control). Tebuconazole, prochloraz and bromuconazole reduced DON concentration by 43%, while epoxiconazole was ineffective. DON concentration in kernels of naturally infected cultivars was 1.95mgkg^–1, a concentration which exceeds the 1mgkg^–1 maximum level of contamination allowed in the United States. Furthermore prochloraz, bromuconazole and tebuconazole applications, in the naturally inoculated trials, reduced DON concentration from 73% to 96%, while epoxiconazole showed the lowest effectiveness. Moreover, a positive linear correlation between Fusarium infected grains and the DON concentration was observed.     

Production of Beauvericin by Different Races of Fusarium Oxysporum F. sp. Melonis, The Fusarium Wilt Agent of Muskmelon

Fourty-four strains of Fusarium oxysporum were isolated from plants of melon with Fusarium wilt symptoms. Among these strains, thirty-nine were characterized for their pathogenicity on melon. Thirty-seven strains belonged to known races of F. oxysporum f. sp. melonis, while two strains were non-pathogenic. Four strains belonged to race 0, seven to race 1, four to race 2, and twenty-two to race 1,2. Beauvericin was produced by thirty-six strains in a range from 1 to 310gg^–1. Eight isolates of race 1,2 did not produce the toxin. In addition, of the two non-pathogenic strains, only one strain produced the toxin (290gg^–1). The production of enniatin A₁, enniatin B₁, and enniatin B was also investigated. Enniatin B was the only enniatin detected, being produced by eleven strains belonging to all the races, with a range of production from traces to 60gg^–1. Finally, melon fruits belonging to two different cultivars (Cantalupo and Amarillo) were artificially inoculated with one strain of F. oxysporum f. sp. melonis (ITEM 3464). Beauvericin was detected in the fruit tissues of both cultivars at a level of 11.2 and 73.8gg^–1, respectively. These data suggest that the production of both the toxins is not related to the pathogenicity of F. oxysporum f. sp. melonis, nor to the differential specificity of the races. The results confirm that beauvericin is a common metabolite of phytopathogenic Fusarium species.     

Scab Response and Moniliformin Accumulation in Kernels of Oat Genotypes Inoculated with Fusarium avenaceum in Poland

Inoculation experiments with 14 genotypes of oats (10 cultivars and 4 lines) were performed during 1996, 1997 and 1998 in Sitaniec, South-Eastern Poland. Panicles of oats were inoculated with a conidial suspension of Fusarium avenaceum, which caused a reduction in yield by 33% and in 1000 kernel weight (TKW) by 21%. During the period between inoculation and harvest, F. avenaceum was able to accumulate moniliformin (MON) in kernels at an average level of 0.13mgkg^–1 (gg^–1). The highest reduction of yield components caused by the F. avenaceum inoculation was found for cv. Santor, followed by lines CHD 1171, STH 2795 and cvs: Kwant and Farys, while cvs Slawko, Dukat, Borys and Komes exhibited the highest resistance to the disease in terms of TKW and yield reductions after inoculation.     

Analysis of gene sequences for the nucleocapsid protein from <Emphasis Type="Italic">Tomato spotted wilt virus</Emphasis> for promoting RNA-mediated cross-protection using the <Emphasis Type="Italic">Potato virus X</Emphasis> vector system

Virus interactions between Tomato spotted wilt virus (TSWV) and Potato virus X (PVX) containing the nucleocapsid protein (N) gene sequences were examined to evaluate the capacity of the N gene sequences from TSWV to promote RNA-mediated cross-protection. Plants simultaneously inoculated with TSWV and PVX containing the 3 96bp of the N gene were highly resistant to TSWV infection, whereas no such resistance was observed in plants inoculated with TSWV and PVX containing the 5 96bp. These results suggest that the 3 portion of the N gene has a higher capacity for promoting RNA-mediated cross-protection of TSWV.     

Induced Resistance to Cyst and Root-knot Nematodes in Cereals by DL-β-amino-n-butyric Acid

Foliar sprays and soil drenches with DL--amino-n-butyric acid (BABA) reduced the number of Heterodera avenae and H. latipons cysts on wheat and barley. Foliar sprays of wheat with 8000mgl^–1 BABA reduced the number of H. avenae cysts by 90%, whereas 2000mgl^–1 BABA was enough to reduce the number of H. latipons cysts by 79%. Multiple spray treatments with 2000mgl^–1 BABA at 10-day intervals reduced the number of H. avenae cysts on wheat and barley. A soil drench of wheat with 125mgl^–1 BABA reduced the number of H. latipons cysts by 93% and H. avenae cysts by 43%. Second-stage juveniles of these nematodes penetrated and formed syncytia in wheat roots soil-drenched with BABA. More adult males of H. avenae were produced in BABA (<250mg1^–1)-treated wheat roots (~76%) than in untreated roots (27%). Soil drenches with higher concentrations of BABA inhibited development of adult males and females. Several chemical elicitors of induced resistance were tested for their ability to reduce the number of H. avenae cysts on wheat. Only BABA was found to be an effective resistance inducer. The number of egg masses of an unidentified Meloidogyne sp. root-knot nematode, which infects only monocots, was also reduced by 95% by a soil drench of wheat with 500mgl^–1 BABA. Development of this nematode inside the BABA-treated roots was also inhibited.     

Electron microscope observations of nuclear polyhedra fromMalacosoma neustria (Lepidoptera: Lasiocampidae

The nuclear polyhedral bodies fromMalacosoma neustria are enclosed within a membrane. The diameter of the nuclear polyhedra varies from 0.9 to 2.8 with an average of 1.8 . In the nuclear polyhedra the rod-like virus particles occur both singly and in bundles. The single virus rods are enclosed within two membranes, namely the intimate membrane and the developmental membrane. The virus rods which occur in bundles have an intimate membrane just like the single virus rods, whereas the developmental membrane encloses the whole bundle. The virus rods are closely packed by the intimate membrane and between the intimate and the developmental membrane is a space. The diameter of the virus rods without membranes, determined from sectioned polyhedra, is about 25 m and the length 250 m.     

Suppressive effect of potassium silicate on powdery mildew of strawberry in hydroponics

From 1996 to 1997, potassium silicate (SiO₂) was tested at 0, 25, 50, and 100mgl^–1 in hydroponics to control powdery mildew. Other elements were added in the usual amounts, and the strawberries were cultivated hydroponically in a greenhouse for 4 months (from October to January). The powdery mildew spread in the control plot, but little mildew developed in the plot with 25mgl^–1 silicate, and none in plots with more than 50mgl^–1 silicate. The suppressive effect lasted for about 4 months on fruits and even longer on leaves. On analysis of mineral content in the leaves, only the silicate content differed markedly between the control and treated plants. Nitrogen, phosphate, potassium, and calcium contents did not differ greatly. The maximum silicate content was about 24 times that of the control, and disease severity decreased significantly when the content was more than 1.5% in the leaves. The hardness of the strawberry leaves, measured by a creep meter, was increased by the silicate treatment.     

Effects of fusaric acid on cells from tomato cultivars resistant or susceptible toFusarium oxysporum f. sp.Lycopersici

Cell suspension cultures were set up from two tomato cultivars, one resistant, (Rio grande) and one susceptible (63.5) toFusarium oxysporum f. sp.lycopersici. Growth rates of the two cell cultures were comparable. Toxicity of fusaric acid, expressed as the fresh weight loss, was analyzed: It was significant in both cases after 10 h, but toxicity was twice as high for 63.5 suspension cells. In the same way, electrolyte leakage caused by fusaric acid was three times more important for 63.5 suspension cells. Moreover, fusaric acid treatment resulted in an acidification of the extracellular medium for 63.5 suspension cells (0.4 pH unit), whereas an alkalization was observed for Rio grande suspension cells (0.2 pH unit). Preliminary experiments suggest that fusaric acid was partially metabolized by Rio grande suspension cells, however, no detoxified forms of fusaric acid were detected either in cells or in culture filtrates. For these two tomato cultivars, the differences in sensitivity to fusaric acid of cultivated cells correspond to the differences in plant susceptibility toFusarium oxysporum f. sp.lycopersici.Abbreviations BAP 6-benzylaminopurine - conductivity - 2,4-D 2,4-dichlorophenoxyacetic acid - EtOAc ethyl acetate - FA fusaric acid - resistivity     

Pathogenic variation in poplar rust <Emphasis Type="Italic">Melampsora larici-populina</Emphasis> from England

Using a leaf disc method, 19 isolates of the poplar rust, Melampsora larici-populina , and one isolate of M.populnea from England were inoculated on to 25 poplar clones belonging to Populus nigra and P.trichocarpa, and hybrids between P. deltoides and P. nigra, P. deltoidesand P. trichocarpa, P.tacamahaca and P.trichocarpa, and P. alba and P. tremula. Disease was scored based on the pustule area and inoculum density. In terms of whether sporulating uredinia formed, the 19 isolates showed seven different patterns to the tested poplar clones. The majority of the rust isolates infected P. nigra P3090 and Vereecken, P.nigra×P. deltoides Casale and Tasman, P. tacamahaca×trichocarpa 36 and Balsam Spire, and P.trichocarpa Blom. Populus trichocarpa×P. deltoides 69039/4 was infected by only three isolates collected from southern England. No visible symptoms appeared on P. alba ×P. tremulaTower and P.trichcarpa×P. deltoides×P. deltoides76028/5 in inoculations with M. larici-populina isolates. Populus alba×P.tremula Tower was infected only by M. populnea. When M. larici-populina isolates were tested using AFLP, no differences were found either between isolates from different geographical regions or between those having narrow spectrum of virulence and those showing wide spectrum of virulence on the tested clones. The results suggest that the UK rust populations possess virulences which were found in races E1, E2, E3 and E4 in continental Europe and that rust having virulence patterns similar to race E4 has occurred in UK poplar plantations since 1996.     

Ribonucleases in the Seedlings of Pearl Millet and their Involvement in Resistance Against Downy Mildew Disease

Tissue homogenates of pearl millet seedlings (cultivars HB 3, 843 B, ICMP 451 and IP 18292), with varying degree of resistance to downy mildew disease were tested for ribonuclease (RNase) enzyme activity and the profile of major RNase isozymes by substrate based gel assay. Polyacrylamide gel electrophoresis (PAGE) of the four pearl millet homogenates revealed 15–20 isozymes, varying in size from 6.5 to 121.0kDa. Most of the RNases were highly active between pH 6 and 8 with maximum activity at pH 7. Tissue specific expression of RNase was observed with more activity in the root, i.e., 38.84, 59.61, 39.90 and 49.23 units in HB 3, 843 B, ICMP 451 and IP 18292, respectively than in shoot 11.54, 9.95, 9.46 and 9.49 units in HB 3, 843 B, ICMP 451 and IP 18292, respectively. Effect of metal ions on the RNase profile indicates Zn⁺⁺ at 2, 20 and 200M concentrations to be inhibitory. Ca⁺⁺ and Mg⁺⁺ at 1mM concentration enhanced the enzyme activity while at 10mM inhibition of enzyme activity was observed. Inoculation with the downy mildew pathogen Sclerospora graminicola reduced RNase activity by 4–13% in compatible interactions while in incompatible combinations, the enzyme activity increased by 10–27%. The significance of RNase in pearl millet-downy mildew interaction and its involvement of in systemic acquired resistance of pearl millet against the downy mildew pathogen are discussed.     

Quantitative studies on dispersal of Pseudocercosporella herpotrichoides spores from infected wheat straw by simulated rain

Splash dispersal ofPseudocercosporella herpotrichoides spores from infected wheat straw was investigated using simulated rainfall (rate 13.8l h^–1 m^–2, volume mean diameter 2.9 mm) and wind (2 m sec^–1) in a raintower/wind tunnel complex. Spores were deposited on the floor of the wind tunnel up to 1 m upwind and 2.5 m downwind from the centre of the straw and impacted on vertical surfaces at heights up to 36 cm above it. Fewer spores were collected with increasing distance from the straw and with increasing height. Most spore-carrying splash droplets were in the size range 400–600 m and very few were less than 200 m.Our results show that these spores are generally dispersed over short distance, which is consistent with field observations.Samenvatting Spetterverspreiding vanPseudocercosporella herpotrichoides van besmet tarwestro werd onderzocht met behulp van nagebootste regen (13,8 l uur^–1 m^–2, gemiddelde druppel diameter 2,9 mm) en wind (snelheid 2 m sec^–1) in een proefopstelling van een regentoren en een windtunnel. Sporen werden op de vloer van de windtunnel gedeponeerd tot 1 m afstand tegen de wind in, gerekend vanaf het centrum van het tarwestro, en met de wind mee tot 2,5 m afstand daarvan. Op verticale vlakken werden sporen op een hoogte van 0 tot 36 cm boven de vloer van de windtunnel opgevangen. Naarmate de afstand en de hoogte toenamen werden er minder sporen gevonden. De meeste spetters met sporen hadden een diameter van 400–600 m en slechts enkele waren kleiner dan 200 m.Onze resultaten tonen aan dat de sporen in het algemeen slechts over korte afstanden verspreid worden, hetgeen overeenkomt met veldwaarnemingen.     

Germination of powdery mildew conidia in vitro on cellulose membranes

Conidia of several powdery mildews were found to germinate to a high and reproducible percentage on 25 m thick cellulose membranes, laid on modified Czapek Dox agar in Petri dishes. Germination on 45 m thick, uncoated cellulose, coated cellulose, cellulose acetate and collodion membranes always was lower and more irregular than on 25 m thick cellulose. Probably, the water permeability of the membranes is of importance in causing a humidity gradient favourable for germination of powdery mildew conidia.Samenvatting Bij incubatie van conidiën van echte meeldauwschimmels op 25 m dikke cellulosemembranen, gelegd op agar in Petri-schalen werden hoge kiemingspercentages verkregen. Kieming op 45 m dikke, ongecoate cellulose, gecoate cellulose, cellulose-acetaat- en collodionmembranen was altijd slechter en onregelmatiger dan op 25 m dikke cellulosemembranen. Waarschijnlijk is de waterdoorlaatbaarheid van de membranen van belang voor de kieming door de vorming van een vochtigheidsgradient.     

Silene armeria,a test plant for carnation etched ring virus

A test plant grown from seed was discovered which may be useful for the detection of carnation etched ring virus (CERV) in carnations. InSilence armeria plants, infected with CERV, necrotic line patterns and blotches may develop about 2–3 weeks after inoculation, but this reaction is not very reliable. WhenS. armeria is infected with carnation mottle virus (CaMV) the plants show hardly perceptible symptoms consisting of a faint mosaic of the leaves and slight stunting of the plants. However, whenS. armeria plants, infected with CaMV, are inoculated with CERV, necrotic lines, rings and blotches develop quite clearly. Thus infection with CaMV makesS. armeria a useful indicator for CERV. Also in the carnation cultivar Joker a synergism exists between the two viruses, severe symptoms resulting from addition of CaMV to CERV-infected plants that otherwise show few if any symptoms.Samenvatting Een uit zaad gekweekte tosplant werd ontdekt, die van belang kan zijn voor het aantonen van het carnation etched ring-virus in anjers. WanneerSilene armeria besmet wordt met dit virus treedt er ongeveer 3 weken na de inoculatie een systemische reactie op bestaande uit necrotische lijnen, kringen en soms vlekken op de bladeren. Deze reactie is evenwel niet erg betrouwbaar. Wordt echter het etched ring-virus geïnoculeerd op planten vanS. armeria die reeds met het carnation mottle-virus besmet zijn — welk virus zwakke symptomen zoals een lichte bontheid in het blad en wat groeivertraging veroorzaakt — dan ontstaan weer de necrotische lijnen, kringen en vlekken. Hierdoor kan de plant als een betrouwbare indicator voor het etched ring — virus worden beschouwd. In de anjercultivar Joker kan ook een dergelijk synergisme aangetoond worden tussen de twee genoemde virussen. Wanneer Joker-planten zonder mottle-virus, maar met etched ring-virus, besmet worden met mottle-virus, verschijnt een heftig ziektebeeld bestaande uit bontheid van het jonge blad en hevige necrotische verschijnselen op de oudere bladeren.     

Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

The isolation of a further new race of Erysiphe graminis DC. f.sp. hordei Marchal and the genetical basis of the resistance of ‘Lyallpur 3645’

Besides the race Amsel-C₂, a new race of barley mildew (Erysiphe graminis DC. f.sp.hordei Marchal) has been isolated from a still resistant variety L 94. This race has provisionally been denoted Lyallpur 3645-C₁₇. It is able to overcome the resistance of Lyallpur 3645.It appears most likely that the barley variety Lyallpur 3645 has two closely linked genes for resistance, one for hypersensitivity and the other for an intermediate type of reaction. The first is present in the variety Amsel and both in Heine 4808.The reactions indicate that race Lyallpur 3645-C₁₇ has only one gene for virulence in addition to the ones present in race Amsel-C₂.A survey is given of the seedling reaction of some 60 previously very or moderately resistant varieties, after infection with the two new races.Samenvatting Twee fysio's van gerstemeeldauw,Erysiphe graminis DC. f.sp.hordei Marchal zijn gevonden, waarvan één niet eerder is beschreven. Dit laatste fysio wordt voorlopig aangeduid als Lyallpur 3645-C₁₇, het andere is het zgn. Amsel-C₂-fysio.Met behulp van beide fysio's is duidelijk geworden dat de lijn Lyallpur-3645 twee nauw gekoppelde resistentiegenen bezit; een gen voor een overgevoeligheidsreactie en een ander gen voor een intermediaire reactie.In het ras Amsel is alleen het overgevoeligheidsgen aanwezig, in Heine 4808, evenals Amsel afkomstig van kruisingen met Lyallpur 3645, zijn beide genen aanwezig.Het fysio Lyallpur-3645-C₁₇ lijkt daarom één gen voor virulentie meer te hebben dan Amsel-C₂.Tenslotte is een overzicht gegeven van de kiemplantreactie na infectie met beide fysio's van ongeveer 60 gerstrassen die eerder zeer goed of matig resistent waren.     

Real-time detection of <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> and <Emphasis Type="Italic">P. citrophthora</Emphasis>in citrus roots and soil

Two primers, specific for Phytophthora nicotianae (Pn6) and P. citrophthora (Pc2B), were modified to obtain Scorpion primers for real-time identification and detection of both pathogens in citrus nursery soils and roots. Multiplex PCR with dual-labelled fluorogenic probes allowed concurrent identification of both species ofPhytophthora among 150 fungal isolates, including 14 species of Phytophthora. Using P. nicotianaespecific primers a delayed and lower fluorescence increase was also obtained from P. cactorumDNA. However, in separate real-time amplifications, the aspecific increase of fluorescence from P. cactorum was avoided by increasing the annealing temperature. In multiplex PCR, with a series of 10-fold DNA dilutions, the detection limit was 10 pg l^-1 for P. nicotianaeand 100 pg l^–1 for P. citrophthora, whereas in separate reaction DNA up to 1 pg l^-1 was detected for both pathogens.Simple and rapid procedures for direct DNA extraction from soil and roots were utilised to yield DNA whose purity and quality was suitable for PCR assays. By combining these protocols with a double amplification (nested Scorpion-PCR) using primers Ph2-ITS4 amplifying DNA from the main Phytophthora species (first round) and PnB5-Pn6 Scorpion and Pc2B Scorpion-Pc7 (second round), it was possible to achieve real-time detection of P. nicotianaeand P. citrophthora from roots and soil. The degree of sensitivity was similar to that of traditional detection methods based on the use of selective media. The analyses of artificially and naturally infested soil showed a high and significant correlation between the concentration of pathogen propagules and the real-time PCR cycle threshold.     

Changes in susceptibility of bean leaves (Phaseolus vulgaris) to Sclerotinia sclerotiorum and Botrytis cinerea by pre-inoculative ozone exposures

The effects of ozone on the susceptibility of leaves ofPhaseolus vulgaris toSclerotinia sclerotiorum andBotrytis cinerea have been investigated. Seedlings of one ozone-sensitive (Pros) and five relatively ozone-insensitive cultivars (Gamin, Precores, Groffy, Narda, Berna) were exposed to different ozone concentrations (0, 120, 180 and 270 g m^–3) for 8 h. One day after the exposures, primary leaves were detached and immediately inoculated with spores of either pathogen suspended in water or in a 62.5 mM KH₂PO₄ (Pi) solution. Visible ozone injury differed between the cultivars and increased with increasing ozone concentration. On the leaves of non-exposed plants, spores of the pathogens suspended in water caused very few lesions, whereas fungal pathogenicity was stimulated by addition of Pi to the inoculum. Ozone-injured leaves of all cultivars exhibited lesions after inoculation of the leaves with the pathogens suspended in water, and the number of lesions was positively correlated with the level of ozone injury for either pathogen and cultivar. The increase in susceptibility of bean leaves in response to increasing ozone concentrations was greater forB. cinerea than forS. sclerotiorum when spores were suspended in water, but was similar when the spores were suspended in Pi.In general, the number of lesions following inoculation with spores in Pi increased with increasing ozone concentration. However, the number of lesions in the ozone-insensitive Groffy was reduced by an exposure to 120 g m^–3 but increased with higher concentrations. This pattern of susceptibility response to the pathogens was not found in the other ozone-insensitive cultivars and, thus, did not appear to be related to the inherent ozone-insensitivity in bean.